Abstract
The method for genetic transformation of maize (Zea mays L.) via embryogenic callus infection with Agrobacterium tumefaciens was developed. Calli were co-cultivated with the overnight culture of A. tumefaciens strain LBA4404 harboring the pBI121 plasmid with the nptII and uidA genes. Thereafter, the sensitivity of calli and regenerated plantlets to kanamycin (Km) was determined. It was shown that kanamycin selection was more efficient at the stage of regenerated plantlets than in callus culture. Both vacuum infiltration at the infection step and preliminary activation of Agrobacterium by acetosyringone or by tobacco leaves exudate increased the frequency of Km-resistant plants. The frequency of Km-resistant plants also varied depending on the morphogenic ability of calli. Polymerase chain reaction confirmed the presence of the nptII gene in the genome of regenerated plants and their progeny. β-Glucuronidase gene expression was observed in roots of T1 plants.
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Abbreviations
- Km:
-
kanamycin
- Cx:
-
cefotaxime
- GUS:
-
β-glucuronidase
- MS me-dium:
-
Murashige and Skoog nutrient medium
- PCR:
-
polymerase chain reaction
REFERENCES
Potrykus, I., Gene Transfer to Cereals: An Assessment, Biotechnology, 1991, vol. 8, pp. 33–37.
Hooykaas van Slogteren, G.M.S., Hooykaas, P.J.J., and Schilperoort, R.A., Expression of Ti-Plasmid Genes in Monocotyledonous Plants Infected with Agrobacterium tumefaciens, Nature, 1984, vol. 311, pp. 763–764.
Schafer, W., Gorz, A., and Kahl, G., T-DNA Integration and Expression in a Monocot Crop Plant after Induction of Agrobacterium, Nature, 1987, vol. 327, pp. 529–532.
Graves, A.C.F. and Goldman, S.L., The Transformation of Zea mays Seedlings with Agrobacterium tumefaciens, Plant. Mol. Biol., 1986, vol. 7, pp. 43–50.
Grimsley, N., Hohn, T., Davis, J.W., and Hohn, B., Agrobacterium Mediated Delivery of Infectious Maize Streak Virus into Maize Plants, Nature, 1987, vol. 325, pp. 177–179.
Gould, J., Devey, M., Hasegawa, O., Eugenio, C.U., Peterson, G., and Smith, R.H., Transformation of Zea mays L. Using Agrobacterium tumefaciens and the Shoot Apex, Plant Physiol, 1991, vol. 95, pp. 426–434.
Ishida, Y., Saito, H., Ohta, S., Hiei, Y., Komari, T., and Kumashiro, T., High Efficiency Transformation of Maize (Zea mays L.) Mediated by Agrobacterium tumefaciens, Nature Biotech., 1996, vol. 14, pp. 745–750.
Lupotto, E., Reali, A., Passera, S., and Chan, M.T., Maize Elite Inbred Lines Are Susceptible to Agrobacterium tumefaciens-Mediated Transformation, Maydica, 1999, vol. 44, pp. 211–218.
Zhao, Z.Y., Gu, W., Cai, T., and Pierce, D.A., Methods for Agrobacterium-Mediated Transformation, US Patent no. 5981 840, Inventors, 1999, no. 9.
Frame, B.R., Shou, H., Chikwamba, R.K., Zhang, Z., Xiang, C., Fonger, T.M., Pegg, S.E.K., Li, B., Nettleton, D.S., Pei, D., and Wang, K., Agrobacterium tumefaciens-Mediated Transformation of Maize Embryos Using a Standard Binary Vector System, Plant Physiol., 2002, vol. 129, pp. 13–22.
Murashige, T. and Skoog, F., A Revised Medium for Rapid Growth and Bioassays with Tobacco Tissue Cultures, Physiol. Plant., 1962, vol. 15, pp. 473–497.
Maniatis, T., Frisch, E.F., and Sambrook, J., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor: Cold Spring Harbor Lab., 1982.
Draper, J., Scott, R., Armitage, P., and Walden, R., Plant Genetic Transformation and Gene Expression: A Laboratory Manual, Oxford: Blackwell Sci., 1988.
Hansen, G. and Chilton, M.D., Agrolistic Transformation of Plant Cell: Integration of T-Strans Generated in Planta, Proc. Natl. Acad. Sci. USA, 1996, vol. 93, pp. 14978–14983.
Danilova, S.A. and Dolgikh, Yu.I., The Stimulatory Effect of the Antibiotic Cefotaxime on Plant Regeneration in Maize Tissue Culture, Fiziol. Rast. (Moscow), 2004, vol. 51, pp. 621–625 (Russ. J. Plant Physiol., Engl. Transl., pp. 559–562).
Danilova, S.A. and Dolgikh, Yu.I., Sposob obrabotki embriogennogo kallusa kukuruzy In Vitro (Method for Treatment of Embryogenic Maize Callus In Vitro), Russia Inventor’s Certificate no. 2 200 760, Byull. Izobret., 2003, no. 8.
Malyshenko, S.I., Tyul’kina, L.G., Zvereva, S.D., and Raldugina, G.N., Transgenic Brassica campestris Plants Expressing the gfp Gene, Fiziol. Rast. (Moscow), 2003, vol. 50, pp. 309–315 (Russ. J. Plant Physiol., Engl. Transl., pp. 276–281).
Dekeyser, R.A., Claes, B., Marichal, M., van Montague, M., and Caplan, A., Evaluation of Selectable Markers for Rice Transformation, Plant Physiol., 1989, vol. 90, pp. 217–223.
Stachel, S.E., Messens, E., van Montagu, M., and Zambryski, P., Identification of the Signal Molecules Produced by Wounded Plant Cells That Activate T-DNA Transfer in Agrobacterium tumefaciens, Nature, 1985, vol. 318, pp. 624–629.
Zakharchenko, N.S., Kalyaeva, M.A., and Bur’yanov, Ya.I., Induction of Agrobacterial T-DNA Processing by Exudates of Monocotyledonous Plants, Fiziol. Rast. (Moscow), 1999, vol. 46, pp. 282–291 (Russ. J. Plant Physiol., Engl. Transl., pp. 239–247).
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Translated from Fiziologiya Rastenii, Vol. 52, No. 4, 2005, pp. 600–607.
Original Russian Text Copyright © 2005 by Danilova, Dolgikh.
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Danilova, S.A., Dolgikh, Y.I. Optimization of Agrobacterial (Agrobacterium tumefaciens) Transformation of Maize Embryogenic Callus. Russ J Plant Physiol 52, 535–541 (2005). https://doi.org/10.1007/s11183-005-0079-5
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DOI: https://doi.org/10.1007/s11183-005-0079-5